Classifications

• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
  • C12N5/10—Cells modified by introduction of foreign genetic material
  • C12N5/12—Fused cells, e.g. hybridomas
  • C12N5/16—Animal cells
  • C12N5/163—Animal cells one of the fusion partners being a B or a T lymphocyte
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
  • C07K16/44—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material not provided for elsewhere, e.g. haptens, metals, DNA, RNA, amino acids
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N1/00—Microorganisms; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
  • C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
  • C12R2001/00—Microorganisms ; Processes using microorganisms
  • C12R2001/91—Cell lines ; Processes using cell lines

Definitions

• the invention relates to new IgE-producing hybrid cell lines, processes for their preparation and their use.
• the invention further relates to a method for producing or obtaining mouse immunoglobulinE (IgE) antibodies with known antigen specificity, with which local and systemic allergic reactions can be generated specifically and reproducibly on experimental animals.
• IgE mouse immunoglobulinE
• IgE antibody with known and reproducible antigen specificity would be just as important for studies on the origin and mechanism of allergic processes as for the detection of anti-allergic substances.
• OA allergen specificity
• DNP known hapten specificity
• the new IgE-antiOA-producing cell line IgE-14-205 was developed by cell fusion between murine myeloma cells and spleen cells from OA-hyperimmunized animals as well as the new IgE-antiDNP-producing cell lines by cell fusion between murine myeloma cells and spleen cells of mice with antibody production against DNP in manufactured in a known manner.
• the new cell lines make it possible for the first time to provide larger amounts of mouse IgE, which has always been associated with considerable difficulties.
• the IgE antibodies produced with the help of the new cell lines are the first IgE antibodies that have a homogeneous antigen specificity and are therefore particularly suitable for triggering standardized allergic reactions.
• they are ideal as an agent for a wide variety of immunochemical, pathophysiological, immunogenetic and pharmacological studies to elucidate the mechanism of allergic reactions.
• experimental animals are understood to mean all animal species that can be used for test purposes.
• Mouse and rat on which the IgE antibodies produced by the method according to the invention in connection with the specific antigen. Ovalbumin can produce reproducible allergic reactions, are particularly suitable for the detection of anti-allergic compounds.
• the highly purified murine IgE from the hybrid cell line IgE-14-205 and from the IgE-antiDNP hybrid cell lines can also be used for the production of anti-mouse IgE antibodies in various animal species.
• the spleen cells are fused with the mycloma cells in the presence of 42% polyethylene glycol for 1-2 minutes at 37 ° (Hämmerling, G.J., Europ. J. Immunol. 7, 743, 1977).
• Hybrid cell lines were obtained by HAT selection (Littlefield, JW, Science 145, 709, 1964), the cell culture supernatants of which were tested 3-5 weeks after cell fusion in the passive cutaneous anaphylaxis test (PCA) for antibody production [PCA for the detection of IgE antiOA antibodies according to Ovary, Z., Immunological Methods, et. J.F.
• the monoclonal hybrid cell line IgE-14-205 was produced according to the illustrated cell hybridization method.
• the PCA titer is given as the reciprocal value of the highest serum or ascites dilution that produces an allergic skin reaction.
• the IgE-14-205 antibody shows the typical heterologous reaction with rat mast cells for mouse immunoglobulin E. It triggers allergic reactions in the rat in connection with the specific allergen, e.g.
• the murine IgE-14-205 antibody produces a passive cutane (PCA) and passive peritoneal anaphylaxis (P P A) in the rat (Tab. 3).
• PCA passive cutane
• P P A passive peritoneal anaphylaxis
• DSCG disodium cromoglycate
• the IgE 14-205 antibody is therefore particularly suitable for testing antiallergic compounds whose potency must be quantitatively reproducibly determined.
• Table 3 shows the inhibition of the passive peritoneal anaphylaxis of the rat triggered by the IgE 14-205 antibody by disodium cromoglycate (Intal R).
• Monoclonal hybrid cell lines with IgE-antiDNP antibody production are produced according to the cell hybridization method shown. '
• the hybrid cell lines permanently synthesize mouse IgE anti-DNP antibodies in the cell culture (e.g. as a suspension culture in DMEM or RPMI 1640 medium), which are secreted into the cell culture medium (Table 1).
• the hybrid cell lines also grow as subcutaneous or intraperitoneal tumors species-specific in Balb / c mice and, under these in vivo conditions, produce large amounts of monoclonal IgE anti-DNP antibodies that can be obtained from the serum and ascites of the tumor-bearing animals ( Tab. 1).
• the IgE-anti-DNP antibodies show the heat instability typical of IgE immunoglobulins: 2 hours at 56 ° C lead to biological inactivation of the antibodies (detection in the PCA test) (Tab. 2).
• the IgE anti-DNP antibodies are hapten specific (Table 2) .DNP, bound to various carrier proteins, e.g. Bovine serum albumin (BSA), or edestin, triggers an allergic reaction (PCA). Amino acids with 2 DNP groups per molecule also trigger an allergic reaction. Amino acids with one DNP group per molecule are not allergenic (bridging theory).
• the IgE anti-DNP antibodies can be precipitated with anti-mouse IgE antiserum.
• the IgE anti-DNP antibodies show the heterologous reaction typical of mouse immunoglobulin E with rat mast cells. They trigger allergic reactions in the rat in connection with the specific hapten (PCA test). Histamine is released from peritoneal rat mast cells, which are sensitized in vitro with IgE-anti-DNP, by anti-mouse IgE antiserum (FIG. 1).
• Fig. 1 Allergic histamine release in vitro
• Peritoneal rat mast cells were sensitized in vitro with various dilutions of IgE à DNP 53-569 ascites. Histamine release was induced by rabbit anti-mouse IgE antiserum (anti-IgE).
• the IgE anti-DNP antibodies are therefore particularly suitable for testing antiallergic compounds, the potency of which must be determined in a reproducible, quantitative manner.
• the IgE anti-DNP antibodies offer the advantage that for the first time animal models for food allergies and percutaneously triggered allergies can be developed.

Landscapes

• Health & Medical Sciences (AREA)
• Chemical & Material Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• Organic Chemistry (AREA)
• Engineering & Computer Science (AREA)
• Genetics & Genomics (AREA)
• Wood Science & Technology (AREA)
• Zoology (AREA)
• Biotechnology (AREA)
• Bioinformatics & Cheminformatics (AREA)
• General Health & Medical Sciences (AREA)
• Biochemistry (AREA)
• General Engineering & Computer Science (AREA)
• Biomedical Technology (AREA)
• Microbiology (AREA)
• Immunology (AREA)
• Medicinal Chemistry (AREA)
• Virology (AREA)
• Tropical Medicine & Parasitology (AREA)
• Mycology (AREA)
• Chemical Kinetics & Catalysis (AREA)
• General Chemical & Material Sciences (AREA)
• Biophysics (AREA)
• Molecular Biology (AREA)
• Proteomics, Peptides & Aminoacids (AREA)
• Cell Biology (AREA)
• Peptides Or Proteins (AREA)
• Preparation Of Compounds By Using Micro-Organisms (AREA)

Priority Applications (1)

Applications Claiming Priority (4)

Publications (3)

Family

ID=25775384

Family Applications (1)

Country Status (2)

Cited By (2)

Citations (2)

• 1979
  • 1979-08-03 EP EP79102797A patent/EP0008092B1/fr not_active Expired
  • 1979-08-03 DE DE7979102797T patent/DE2963410D1/de not_active Expired

Patent Citations (2)

Non-Patent Citations (7)

Also Published As

Similar Documents

Legal Events